Permeation kinetics of active drugs through lanolin-based artificial membranes.

Skin-penetration studies play an essential role in the selection of drugs for dermal or transdermal application. In vivo experiments in humans are not always possible for ethical, practical, or economic reasons, especially in the first part of the drug development. It is necessary to develop alternative methods using accessible and reproducible surrogates for in vivo human skin. The in vitro methodologies using biological membranes (human and animal skin) are recognized and well accepted as an alternative but present high inter- and intra-individual variability. Therefore, the formation of synthetic membranes has been studied to obtain skin- mimicking models for permeation studies. The aim of this work is to create lanolin-based artificial membranes that can mimic the absorption through the skin of compounds applied topically. A series of synthetic membranes using two different types of lanolin (water-extracted (WE) and solvent-extracted (SE)) were prepared. Next, the in vitro release test of three drugs (diclofenac sodium, ibuprofen and lidocaine) was performed on artificial membranes and on porcine skin placed on Franz cells. The percentage of release, flux, permeability coefficient, lag time, area under the curve, maximal concentration and time were determined for each compound in the different types of membrane. The results showed that lanolin membranes presented a strong diminution of permeability compared to most artificial membranes, leading to a very similar permeability to that of skin. The SE and WE membranes showed a diminution of transepidermal water loss and permeability of compounds compared with membranes alone. The results from WE membranes were similar to those found for the skin. The lanolin membranes were not capable of perfectly mimicking permeation through the skin, but they did have the same rank order of drug penetration as the skin. It may be deduced from these tests that these systems provide more reliable results for compounds with low to medium lipophilicity. The results demonstrated that new lanolin-based artificial membranes have the potential to be exploited as screening models for determining the permeability of a compound destined to be topically delivered.

Caffeine delivery in porcine skin: a confocal Raman study.

Confocal Raman microscopy is a novel optical method for studies of pro-drug and drug delivery. This method is a promising technique that enables non-destructive measurement of the permeation profile through skin layers. Peaks of compounds are usually normalised to skin peaks (amino-acid and amide I) for semi-quantitative evaluation. The present study seeks to optimise a methodology for complete quantitative measurement of the amount of an active compound at different depths. Caffeine was used as a tracer to evaluate compound's skin penetration using confocal Raman microscopy. A semi-quantitative depth profile of caffeine was obtained with normalisation of the Raman intensities. These ratios of Raman intensities were correlated with the caffeine concentration using an external calibration curve. The calibration curve was carried out with porcine skin incubated in different concentrations of caffeine; afterwards, each skin sample was analysed by confocal Raman microscopy and HPLC to determine the relation between the Raman signal intensity and the caffeine concentration per skin mass and to create a depth profile. These correlation curves allow the full quantification of the caffeine in skin from Raman intensity ratios at different depths.

Skin permeation and antioxidant efficacy of topically applied resveratrol.

The permeation of resveratrol was assessed by in vitro and in vivo experiments 24 h after topical administration. The in vitro profile of resveratrol was assessed by Raman spectroscopy. Human skin permeation was analysed in vivo by the tape stripping method with the progressive removal of the stratum corneum layers using adhesive tape strips. Moreover, the free radical scavenging activity of resveratrol after its topical application was determined using the DPPH assay. The Raman spectra indicated that the topically applied resveratrol penetrates deep into the skin. The results showed high amounts of resveratrol in the different stratum corneum layers close to the surface and a constant lower amount in the upper layers of the viable epidermis. The concentration of resveratrol present in the outermost stratum corneum layers was obtained by tape stripping after in vivo application. The results demonstrated that resveratrol mainly remained in the human stratum corneum layers. After topical application, resveratrol maintained its antiradical activity. The antioxidant efficacy of the compound was higher in the inner layers of the stratum corneum. As these results have demonstrated, topically applied resveratrol reinforces the antioxidant system of the stratum corneum and provides an efficient means of increasing the tissue levels of antioxidants in the human epidermis.

Exogenous and endogenous lipids of human hair.

PURPOSE: The aim of this study was to characterize the external and internal lipids of Caucasian hair and their influence in different hair properties such as moisture content, hydrophobic character, and mechanical properties. METHODS: Lipid extraction and their analysis by thin layer chromatography with flame ionization detector were carried out. Lipid rearrangement and water sorption and desorption evaluation of these fibers with and without lipids will also be determined using different techniques such as differential scanning calorimetry, thermogravimetric analysis and dynamic vapor sorption, mainly to evaluate permeation changes of these hair fibers possibly related to the fluidity of the lipids extracted. RESULTS: Caucasian fibers were found to be well hydrated, and moisture diminution was observed mainly for the external lipid extracted fibers. Unexpectedly, the lipid extraction promoted an increase in the break tenacity of the Caucasian fibers. The hydrophobic character of the fiber surfaces indicates the marked hydrophobicity of all fibers. Delipidization promotes only a slight diminution of their hydrophobic properties. Water uptake and desorption studies indicate an important water regain for Caucasian fibers. The external extracted hair fibers presented a diminution of maximum water regain, which surprisingly increases with the following internal lipid extraction. This can be due to a higher water desorption found only for external extracted fibers. CONCLUSIONS: The relationship between fluidity of lipids extracted and hair fiber water diffusion were established. Extraction of internal lipids of Caucasian fibers, which have a higher unsaturated lipid content than external lipids of the same hair fiber, leads to a lower water permeability of the fiber. On the capillar formulations should be considered the importance of lipid fluidity to modify the permeability of the fiber.

BRG1/SMARCA4 is essential for neuroblastoma cell viability through modulation of cell death and survival pathways.

Neuroblastoma (NB) is a neoplasm of the sympathetic nervous system, and is the most common solid tumor of infancy. NBs are very heterogeneous, with a clinical course ranging from spontaneous regression to resistance to all current forms of treatment. High-risk patients need intense chemotherapy, and only 30-40% will be cured. Relapsed or metastatic tumors acquire multi-drug resistance, raising the need for alternative treatments. Owing to the diverse mechanisms that are responsible of NB chemoresistance, we aimed to target epigenetic factors that control multiple pathways to bypass therapy resistance. We found that the SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4 (SMARCA4/BRG1) was consistently upregulated in advanced stages of NB, with high BRG1 levels being indicative of poor outcome. Loss-of-function experiments in vitro and in vivo showed that BRG1 is essential for the proliferation of NB cells. Furthermore, whole-genome transcriptome analysis revealed that BRG1 controls the expression of key elements of oncogenic pathways such as PI3K/AKT and BCL2, which offers a promising new combination therapy for high-risk NB.

Skin penetration and antioxidant effect of cosmeto-textiles with gallic acid.

In this work, the antioxidant gallic acid (GA) has been encapsulated in microspheres prepared with poly-ε-caprolactone (PCL) and incorporated into polyamide (PA) obtaining the cosmeto-textile. The topical application of the cosmeto-textile provides a reservoir effect in the skin delivery of GA. The close contact of the cosmeto-textile, containing microsphere-encapsulated GA (ME-GA), with the skin and their corresponding occlusion, may be the main reasons that explain the crossing of active principle (GA) through the skin barrier, located in the stratum corneum, and its penetration into the different compartments of the skin, epidermis and dermis. An ex vivo assessment was performed to evaluate the antioxidant effect of the ME-GA on the stratum corneum (SC) using the thiobarbituric acid-reactive species (TBARS) test. The test is based on a non-invasive ex vivo methodology that evaluates lipid peroxides formed in the outermost layers of the SC from human volunteers after UV radiation to determine the effectiveness of an antioxidant. In this case, a ME-GA cosmeto-textile or ME-GA formulation were applied to the skin in vivo and lipid peroxidation (LPO) in the horny layer were determined after UV irradiation. This methodology may be used as a quality control tool to determine ex vivo the percentage of LPO inhibition on human SC for a variety of antioxidants that are topically applied, in this case GA. Results show that LPO formation was inhibited in human SC when GA was applied directly or embedded in the cosmeto-textile, demonstrating the effectiveness of both applications. The percentage of LPO inhibition obtained after both topical applications was approximately 10% for the cosmeto-textile and 41% for the direct application of microspheres containing GA. This methodology could be used to determine the effectiveness of topically applied antioxidants encapsulated in cosmeto-textiles on human SC.

The influence of hair lipids in ethnic hair properties.

OBJECTIVE: Biochemical studies have mainly focused on the composition of hair. African hair exhibited lower moisturization and less radial swelling when flushing with water compared with Asian or Caucasian hair, and they assumed a possible lipid differentiation among human populations. This study consists in the lipid characterization of different ethnic hairs (Caucasian, Asian and African hairs) and the influence of these lipids in different hair properties such as humidity and mechanical properties. Evaluation of water sorption and desorption of the different ethnic hairs and with and without lipids is also studied mainly to determine permeation changes of the keratin fibres. METHODS: Extractions of exogenous and endogenous lipids with different organic solvents were performed; lipid analysis and its quantification using thin-layer chromatography coupled to an automated flame ionization detector (TLC/FID) were performed. Absorption and desorption curves were obtained in a thermogravimetric balance equipped with a controlled humidity chamber, the Q5000SA Sorption Analyzer (TA Instruments, New Castle, IL, U.S.A.). Also, mechanical properties (breaking stress and breaking elongation) were analysed using a computer programmable dynamometer (Instron 5500R). RESULTS: Lipid extraction showed the highest amount of total lipids for the African hair which may come from external sebaceous lipids compared with Asian or Caucasian hair. Caucasian fibres were found to be the most hydrated fibre, and a decrease in moisture was found in the extracted fibres, again, which is more important for the Caucasian hair. A superior lineal mass was found for the Asian fibres which supported their higher strength. The results obtained from the analysis of the mechanical properties of delipidized fibres indicate a surprising increase in the strength of African and Caucasian fibres. Perhaps this increase in strength could be related to the humidity decrease in lipid-extracted hair fibres. Results of water uptake and desorption indicate that Asian and Caucasian hairs present the lower diffusion coefficients compared with the African ones. At least for the African fibre, an extraction of its lipids that mainly account for apolar lipids ameliorates the fibre structure, decreasing its permeability to water and increasing its tensile strength. CONCLUSION: The ethnic hairs were assessed related to their lipid composition, and some differences between them were found in terms of water uptake and mechanical properties.

Advanced hair damage model from ultra-violet radiation in the presence of copper.

OBJECTIVE: Damage to hair from UV exposure has been well reported in the literature and is known to be a highly complex process involving initiation via absorption of UV light followed by formation and propagation of reactive oxygen species (ROS). The objective of this work was to understand these mechanisms, explain the role of copper in accelerating the formation of ROS and identify strategies to reduce the hair damage caused by these reactive species. METHODS: The location of copper in hair was measured by Transmission electron microscopy-(TEM) X-ray energy dispersive spectroscopy (XEDS) and levels measured by ICP-OES. Protein changes were measured as total protein loss via the Lowry assay, and MALDI ToF was used to identify the biomarker protein fragments. TBARS assay was used to measure lipid peroxide formation. Sensory methods and dry combing friction were used to measure hair damage due to copper and UV exposure and to demonstrate the efficacy of N,N' ethylenediamine disuccinic acid (EDDS) and histidine chelants to reduce this damage. RESULTS: In this work, a biomarker protein fragment formed during UV exposure is identified using mass spectrometry. This fragment originates from the calcium-binding protein S100A3. Also shown is the accelerated formation of this peptide fragment in hair containing low levels of copper absorbed from hair during washing with tap water containing copper ions. Transmission electron microscopy (TEM) X-ray energy dispersive spectroscopy (XEDS) studies indicate copper is located in the sulphur-poor endo-cuticle region, a region where the S100A3 protein is concentrated. A mechanism for formation of this peptide fragment is proposed in addition to the possible role of lipids in UV oxidation. A shampoo and conditioner containing chelants (EDDS in shampoo and histidine in conditioner) is shown to reduce copper uptake from tap water and reduce protein loss and formation of S100A3 protein fragment. In addition, the long-term consequences of UV oxidation and additional damage induced by copper are illustrated in a four-month wear study where hair was treated with a consumer relevant protocol of hair colouring treatments, UV exposure and regular shampoo and conditioning. CONCLUSIONS: The role of copper in accelerating UV damage to hair has been demonstrated as well as the ability of chelants such as EDDS and histidine in shampoo and conditioner products to reduce this damage.

In vivo and in vitro evaluation of topical formulations containing physiological lipid mixture for replacement of skin barrier function.

AIM: The aim of the study was to describe a new in vivo and in vitro approach of the efficacy evaluation of cosmetic emollients to better understand the link between the formulation and the activity of cosmetic products. METHODS: Two long term in vivo studies were carried out on nine healthy Caucasian volunteers mean age 40±12 years to evaluate the protecting and repairing effects of the two different barrier repair cosmetic formulations. The application of the formulations was repeated once a day during 7 days and biophysical parameters (TEWL and Skin Hydration) were measured before and after Sodium laureth sulphate exposure The in vitro study was carried out by freeze substitution transmission electron microscopy (FSTEM) on stratum corneum samples obtained by sections of fresh skin from young pigs, depleted with a solvent mixture and treated with the two products RESULTS: The in vivo results demonstrated that daily product application provided a reinforcement of the skin barrier with protecting and repairing effects from chemical injuries the extent of which was dependent on the formulation features (product A>product B) The role of the technical form on the lipid availability was confirmed by the in vitro evaluation tests. CONCLUSION: The results point out that a daily application of physiological lipid mixture containing emulsion can protect healthy skin and promote the reparing effect on unpaired barrier skin, reducing TEWL and maintaining hydration of the stratum corneum. The efficacy degree is higher when the cosmetic form promotes the availability of active ingredients increasing the product performance.

Bicellar systems as vehicle for the treatment of impaired skin.

This study assesses the potential usefulness of bicellar systems to retard the penetration of drugs into damaged skin. The active compound used in this study was diclofenac diethylamine (DDEA). Initially, physicochemical characterisation of the DDEA bicellar systems was performed at different temperatures by small-angle X-ray scattering (SAXS), wide-angle X-ray scattering (WAXS) and differential scanning calorimetry (DSC) techniques. Subsequently, in vitro percutaneous absorption of bicellar systems into in vitro damaged skin was studied. SAXS results indicated a slight decrease in the width of their bilayers with increasing temperature, with no apparent stacking in those systems. WAXS patterns were compatible with an orthorhombic lateral packing of the nanoaggregates. The thermogram obtained by DSC indicated a decrease in gel-to-liquid crystalline transition temperature (Tm) when the drug was included into bicellar systems. A retardation effect for DDEA was detected by in vitro percutaneous absorption studies when DDEA was vehiculised in the bicellar systems with respect to an aqueous solution of the drug. It seems that the use of bicellar systems as a vehicle for topical application of DDEA on skin with an impaired barrier function may inhibit the penetration of DDEA to the systemic level. Such systems may consequently repair stratum corneum barrier function to some extent. The use of these systems could be considered a new alternative strategy to treat topically pathological skin with different drugs.

Bicellar systems as new delivery strategy for topical application of flufenamic acid.

In this work, bicellar systems, bilayered disc-shaped nanoaggregates formed in water by phospholipids, are proposed as a novel strategy for delivery of the anti-inflammatory flufenamic acid (FFA) to the skin. A comparative percutaneous penetration study of this drug in bicellar systems and other vehicles was conducted. The effects induced on the skin by the application of FFA in the different vehicles were analyzed by attenuated total reflectance-fourier transform infrared (ATR-FTIR). Additionally, using the microscopic technique freeze-substitution transmission electron microscopy (FSTEM) and X-ray scattering technique using synchrotron radiation (SAXS-SR), we studied the possible microstructural and organizational changes that were induced in the stratum corneum (SC) lipids and the collagen of the skin by the application of FFA bicellar systems. Bicellar systems exhibited a retarder effect on the percutaneous absorption of FFA with respect to the other vehicles without promoting disruption in the SC barrier function of the skin. Given that skin disruption is one of the main effects caused by inflammation, prevention of disruption and repair of the skin microstructure should be prioritized in anti-inflammatory formulations.

Keratins and lipids in ethnic hair.

Human hair has an important and undeniable relevance in society due to its important role in visual appearance and social communication. Hair is mainly composed of structural proteins, mainly keratin and keratin associated proteins and lipids. Herein, we report a comprehensive study of the content and distribution of the lipids among ethnic hair, African, Asian and Caucasian hair. More interestingly, we also report the study of the interaction between those two main components of hair, specifically, the influence of the hair internal lipids in the structure of the hair keratin. This was achieved by the use of a complete set of analytical tools, such as thin layer chromatography-flame ionization detector, X-ray analysis, molecular dynamics simulation and confocal microscopy. The experimental results indicated different amounts of lipids on ethnic hair compositions and higher percentage of hair internal lipids in African hair. In this type of hair, the axial diffraction of keratin was not observed in X-ray analysis, but after hair lipids removal, the keratin returned to its typical packing arrangement. In molecular dynamic simulation, lipids were shown to intercalate dimers of keratin, changing its structure. From those results, we assume that keratin structure may be influenced by higher concentration of lipids in African hair.

Skin efficacy of liposomes composed of internal wool lipids rich in ceramides.

Ceramides from intercellular lipids of skin stratum corneum are known to play an essential role in maintaining and structuring the lipid barrier of the skin. Internal wool lipids (IWL), which are also rich in ceramides, have a composition similar to that of the stratum corneum lipids. IWL extracted with chloroform/methanol azeotrope at the laboratory scale have been shown to be capable of forming liposomes with a stable bilayer structure. Furthermore, topical application of these IWL liposomes on intact and compromised skin has been demonstrated to improve barrier skin properties.In this study we evaluated the effect on human skin repair of different IWL extract compositions obtained by two extraction methodologies. The formation and characteristics of the liposomes prepared were greatly influenced by the IWL composition, primarily the sterol sulfate content. The IWL liposomes improved skin barrier integrity and increased skin hydration when applied onto intact skin. These improvements were slightly enhanced in the case of IWL liposomes that were richer in polar lipids.

Bicellar systems for in vitro percutaneous absorption of diclofenac.

This work evaluates the effect of different bicellar systems on the percutaneous absorption of diclofenac diethylamine (DDEA) using two different approaches. In the first case, the drug was included in bicellar systems, which were applied on the skin and, in the second case, the skin was treated by applying bicellar systems without drug before to the application of a DDEA aqueous solution. The characterization of bicellar systems showed that the particle size decreased when DDEA was encapsulated. Percutaneous absorption studies demonstrated a lower penetration of DDEA when the drug was included in bicellar systems than when the drug was applied in an aqueous solution. This effect was possibly due to a certain rigidity of the bicellar systems caused by the incorporation of DDEA. The absorption of DDEA on skin pretreated with bicelles increased compared to the absorption of DDEA on intact skin. Bicelles without DDEA could cause certain disorganization of the SC barrier function, thereby facilitating the percutaneous penetration of DDEA subsequently applied. Thus, depending on their physicochemical parameters and on the application conditions, these systems have potential enhancement or retardant effects on percutaneous absorption that result in an interesting strategy, which may be used in future drug delivery applications.

Liposome formation with wool lipid extracts rich in ceramides.

Internal wool lipids (IWLs) are rich in cholesterol, free fatty acids, cholesteryl sulfate, and, mainly, ceramides. The repairing effect of these lipids structured as liposomes was demonstrated by reinforcing the skin-barrier integrity and increasing the water-holding capacity when applied onto the skin. This work was focused on the formation of liposomes with IWLs rich in ceramides, obtained at pilot plant level with organic solvent extraction by using methanol and acetone. The lipid composition of the two extracts was quantitatively analyzed. IWL extracts containing different amounts of sterol sulfate were used to form liposomes at physiologic p(H). Vesicle size distribution, polydispersity index, and zeta potential of all liposomes were determined to characterize them and to study their stability. The results obtained showed that IWL extract composition, which was different depending on the extraction methodologies used, greatly influences the characteristics of the liposomes formed. Vesicular size and polydispersity index liposomes were smaller when the extract composition contained a higher proportion of either free fatty acids or sterol sulfate. Moreover, liposome stability was improved when some amount of sterol sulfate was added to the composition of methanol and acetone extracts. This natural mixture with keratinaceous origin could have a special interest for cosmetic or dermopharmaceutical companies.

Lipid nanostructures: self-assembly and effect on skin properties.

This work evaluates the relation between the composition and the self-assembly of some lipid aggregates with their effects on the skin. To this end, liposomes, bicelles and micelles formed by dipalmitoylphosphatidylcholine (DPPC), dimyristoylphosphatidylcholine (DMPC) and dihexanoylphosphatidylcholine (DHPC) were characterized by electron microscopy and dynamic light scattering techniques, and applied on the skin. The results revealed that nanostructures with similar assembly but different composition caused different effects on the skin parameters. In general, samples containing DMPC affected the barrier function to a greater extent than systems containing DPPC. Additionally, our results showed that samples with the same lipid composition but different assembly exerted different effects on the skin. Liposomes decreased or did not modify the transepidermal water loss (TEWL), while bicelles and micelles increased this parameter. Hydration of the skin diminished especially after the application of micellar and bicellar samples. In vitro experiments showed structures like vesicles inside cutaneous SC (stratum corneum) incubated with DPPC/DHPC bicelles. These structures were not detected in SC samples incubated with DMPC/DHPC bicelles probably due to the different thermotropic behavior of DMPC and DPPC at physiological temperatures. Results reported in this work should be considered in terms of design of more efficient and specific skin delivery systems.

Lamellar rearrangement of internal lipids from human hair.

The internal lipids were extracted from untreated hair without surface lipids. Liposomes were formed with the internal lipids at different hydration levels to determine the organization of these lipids and the influence of the water content on the lamellar structure of the hair fibres by X-ray Scattering (SAXS). Two structures of hair lipids were observed at 4.5 and approximately 9.0 nm with a different behaviour as a function of water content: the largest bilayer being the one that showed a capacity to retain water inside its structure. SAXS was also applied directly to three samples: a packed swatch of hair fibres at 60% RH, fibres soaked in water and delipidized fibres. Only the lamella at 9.0 nm was slightly affected by water content. Moreover, there was a small diminution in intensity probably due to a high permeability of wet fibres which could give rise to a disorder of the lipid structure. These two lamellar rearrangements are probably made up of lipids with a different and specific hydrophilic/hydrophobic balance.

Cosmetic effectiveness of topically applied hydrolysed keratin peptides and lipids derived from wool.

BACKGROUND/PURPOSE: Skin moisturisation, elasticity, feel and appearance can all be improved through the topical application of protein hydrolysates. Recent studies suggest that supplementing intercellular lipids of the stratum corneum can enhance the functioning of the skin. METHODS: In this study, a hydrolysed keratin peptide (molecular weight <1000 Da) was prepared from wool and tested on skin in two different formulations: an aqueous solution and an internal wool lipids (IWL) liposome suspension. In vivo long-term studies were performed to evaluate the water barrier function of the skin after topical application of different formulations. During the treatment period, hydration and elasticity were determined. A sorption-desorption test was also performed to assess the hygroscopic properties and water-holding capacity of the different treated skin sites. RESULTS: Significant differences were found between the control and treated sites, with the treated areas showing an increase in hydration and elasticity as a result of keratin peptide application. Measurements also indicated that the keratin formulations reinforce the skin barrier integrity, improving its water-holding capacity. CONCLUSION: A combination of the keratin peptide with the IWL showed beneficial effects, indicating that this combination is suitable for designing new cosmetics products.

Effect of bicellar systems on skin properties.

Bicelles are discoidal aggregates formed by a flat dimyristoyl-glycero-phosphocholine (DMPC) bilayer, stabilized by a rim of dihexanoyl-glycero-phosphocholine (DHPC) in water. Given the structure, composition and the dimensions of these aggregates around 10-50 nm diameter, their use for topical applications is a promising strategy. This work evaluates the effect of DMPC/DHPC bicelles with molar ratio (2/1) on intact skin. Biophysical properties of the skin, such as transepidermal water loss (TEWL), elasticity, skin capacitance and irritation were measured in healthy skin in vivo. To study the effect of the bicellar systems on the microstructure of the stratum corneum (SC) in vitro, pieces of native tissue were treated with the aforementioned bicellar system and evaluated by freeze substitution applied to transmission electron microscopy (FSTEM). Our results show that bicelles increase the TEWL, the skin elastic parameters and, decrease skin hydration without promoting local signs of irritation and without affecting the SC lipid microstructure. Thus, a permeabilizing effect of bicelles on the skin takes place possibly due to the changes in the phase behaviour of the SC lipids by effect of phospholipids from bicelles.

Wool peptide derivatives for hand care.

Hands experience much greater wear and tear during normal daily routines compared with most other parts of the body, and thereby demand specific needs from cosmetics targeted at hand care. Keratin proteins are the major structural component of the outer layers of the skin. In this work a novel keratin fraction from wool, which has high cystine content present in the S-sulphonated form, has been developed to target hand care applications. In vivo long-term studies were performed to evaluate the water-holding capacity and elasticity of hand skin following topical application of keratins. Moreover, protection of healthy skin against detergent-induced dermatitis was evaluated after topical application of the keratin-active formulation. Significant results in the measured biophysical parameters were found, which indicated an improvement in the skin's water-holding capacity, hydration, and elasticity for volunteers with dry skin as a result of the keratin peptide treatment. Results also indicated that the keratin peptide treatment can prevent some of the damaging effects associated with surfactant exposure.